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Abstract: FR-PO0671

Suppression of ADPKD Alters Epithelial Cell Transcription Program

Session Information

Category: Genetic Diseases of the Kidneys

  • 1201 Genetic Diseases of the Kidneys: Monogenic Kidney Diseases

Authors

  • Rai, Victoria, Yale School of Medicine, New Haven, Connecticut, United States
  • Failli, Mario, Telethon Institute of Genetics and Medicine, Pozzuoli, Campania, Italy
  • Fiusco, Maurizio, Telethon Institute of Genetics and Medicine, Pozzuoli, Campania, Italy
  • Reyna-Neyra, Marcela Andrea, Yale School of Medicine, New Haven, Connecticut, United States
  • Sorrentino, Nicolina Cristina, Telethon Institute of Genetics and Medicine, Pozzuoli, Campania, Italy
  • Onuchic, Laura, Yale School of Medicine, New Haven, Connecticut, United States
  • Di Bernardo, Diego, Telethon Institute of Genetics and Medicine, Pozzuoli, Campania, Italy
  • Craft, Joseph E., Yale School of Medicine, New Haven, Connecticut, United States
  • Caplan, Michael J., Yale School of Medicine, New Haven, Connecticut, United States
Background

Expression of the 200 amino acid C-terminal tail (CTT) portion of the polycystin-1 protein, encoded by the Pkd1 gene whose loss causes ADPKD, suppresses cystic phenotype in late-stage disease in mice. To identify transcriptional differences and to define differential cell states potentially associated with cystogenesis, we performed single cell (scRNA-seq) and bulk RNA sequencing on kidneys from a CTT-suppressed and non-suppressed ADPKD model at an early stage of disease.

Methods

Orthologous doxycycline-inducible mouse models of ADPKD (Pkd1fl/fl;Pax8rtTA;Teto-Cre;C57BL/6N) expressing CTT (Pkd1-KO+CTT) or not (Pkd1-KO) were induced between 4-6 weeks of age and aged to 10 weeks. scRNA-seq was performed on single cell kidney suspensions from biological replicates that were pooled and processed with 10X Genomics. Bulk RNA-seq was performed on whole kidney RNA from biological replicates.

Results

scRNA-seq identified clusters in Pkd1-KO that expressed markers from multiple tubule segments and novel transcripts, indicating that loss of Pkd1 may result in dedifferentiated cells. These clusters are nearly absent in Pkd1-KO+CTT. Pseudotime and RNA velocity suggest CTT expression maintains cells in an intermediate state, preventing them from reaching the Pkd1-KO associated clusters which represent a more advanced state. Bulk RNA-seq further revealed genes that are not only differentially expressed between Pkd1-KO and Pkd1-KO+CTT, but also genes that are differentially expressed between each model and non-cystic wildtype kidneys. Disease endpoints are not significantly different at this early stage of disease between Pkd1-KO and Pkd1-KO+CTT, supporting a cell-autonomous CTT influence on epithelial cell transcription program. Ongoing work investigates the non-cell-autonomous features of CTT expression, including its influence on immune cell recruitment and its potential suppression of Pkd2-related cystic disease.

Conclusion

Transcriptomic results reveal that CTT expression alters epithelial cell transcription program in early ADPKD, which may contribute to disease suppression.

Funding

  • NIDDK Support

Digital Object Identifier (DOI)